Multi-omics peripheral nerve atlas enables fine-mapping of pain molecular phenotypes

多组学周围神经图谱能够精细绘制疼痛分子表型

基本信息

批准号：
10707409
负责人：
JEFFREY D MILBRANDT
金额：
$ 72.78万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707409
关键词：
Address American Atlases Axon Candidate Disease Gene Cell Nucleus Cells Chromatin Clinical Communities Cutaneous Data Data Set Disease Failure Femur Foundations Functional disorder Gene Combinations Gene Expression Gene Expression Profile Genes Genetic Transcription Genome Genomic approach Genomics Genotype Goals Grain Human Immune Knowledge Lateral Macrophage Maps Medical Methods Molecular Motor Mus Mutation Nerve Neuroglia Neurologic Neuroma Neurons Nociceptors Ontology Operative Surgical Procedures Organ Donor Pain Pain Map Pathway Analysis Pathway interactions Patients Pattern Peripheral Nerves Peripheral Nervous System Phenotype Pre-Clinical Model Proteomics RNA Research Resected Resolution Resources Rodent Sampling Schwann Cells Sensory Series Statistical Data Interpretation Syndrome Technology Therapeutic Tissue Donors Tissue Procurements Tissues Traumatic injury Variant XCL1 gene abuse liability afferent nerve axonal sprouting candidate validation cell type chronic pain chronic painful condition cohort data integration experimental study gene discovery genetic variant genome analysis genome sequencing insight molecular phenotype multiple omics neuron component novel novel therapeutics opioid overuse pain model painful neuropathy patient stratification peripheral pain pre-clinical response risk variant single nucleus RNA-sequencing transcriptome transcriptome sequencing transcriptomics transmission process whole genome

项目摘要

Project Abstract – Project 1 Chronic pain afflicts over 100 million Americans and is a leading reason for seeking medical treatment. Few new therapies have been delivered in the past decades, resulting in overuse of opioids, despite their potential for abuse and limited efficacy in many chronic pain conditions. One glaring concern is the failure of studies using preclinical models to yield agents that are effective pain therapeutics. A key barrier in addressing this problem is the knowledge gap between our understanding of the human peripheral nervous system and the studies performed in rodent preclinical pain models. In this project, we propose to identify additional genes involved in human pain pathways via analysis of the genomes of a cohort of patients with idiopathic painful neuropathy. While nociceptive neurons are the components most critical for peripheral pain transmission, other cells in peripheral nerves—glia and immune cells, particularly macrophages—are important regulators of pain responses in these neurons. Additional analysis is required to understand how interactions among these cellular nerve components, and with sensory axons, contribute to pain. We therefore propose to use single-nuclei RNAseq strategies that we recently utilized to produce a mouse peripheral nerve atlas to create a multi-layered, comprehensive atlas of the expression profiles of human peripheral nerve cellular components. In this proposal, we outline a series of experiments to examine gene expression in cells of human peripheral nerve and to identify novel gene variants associated with idiopathic painful neuropathy. Whole genome sequencing will be performed on samples from a large cohort of well-characterized patients with painful neuropathy. Gene variants will be associated with phenotypic information to identify new genes involved in the pain pathway. Using multi-omic technologies, including snRNA-seq and snATAC-seq as well as spatial transcriptomics and multiplex proteomics, to study nerves from our donor tissue core, we will devise a comprehensive, unbiased single-nuclei atlas of gene expression and cellular composition of human peripheral nerves. Parallel studies will be performed to determine the cellular components and gene expression patterns of surgically resected painful neuromas. By comparing the neuroma transcriptomes to the atlas derived from normal nerve datasets, we plan to identify cell type-specific determinants of painful neuromas. We expect these rich and complementary datasets to provide a foundation for molecular and cellular analysis of sensory nerve dysfunction in pain syndromes that will provide insights into treatment options for pain.

项目摘要 – 项目 1 慢性疼痛困扰着超过一亿美国人，并且是寻求医疗治疗的主要原因。过去几十年来提供的治疗方法导致了阿片类药物的过度使用，尽管它们具有潜在的治疗作用滥用和在许多慢性疼痛病症中疗效有限是一个明显的担忧是使用该药物的研究失败。产生有效疼痛治疗药物的临床前模型是解决这一问题的一个关键障碍。我们对人类周围神经系统的理解与研究之间的知识差距在该项目中，我们建议鉴定参与啮齿类动物临床前疼痛模型的其他基因。通过分析一组特发性疼痛性神经病患者的基因组来研究人类疼痛通路。虽然伤害性神经元是外周疼痛传递最关键的成分，但其他细胞周围神经——神经胶质细胞和免疫细胞，特别是巨噬细胞——是疼痛的重要调节因子需要进行额外的分析来了解这些细胞之间的相互作用。神经成分和感觉轴突会导致疼痛，因此我们建议使用单核。我们最近使用 RNAseq 策略来生成小鼠周围神经图谱，以创建多层、人类周围神经细胞成分表达谱的综合图谱。在本提案中，我们概述了一系列实验来检查人类细胞中的基因表达周围神经并识别与特发性疼痛性神经病相关的新基因变异。将对来自一大群特征明确的患有疼痛的患者的样本进行基因组测序基因变异将与表型信息相关联，以识别参与神经病变的新基因。使用多组学技术，包括 snRNA-seq 和 snATAC-seq 以及空间转录组学和多重蛋白质组学，为了研究来自我们供体组织核心的神经，我们将设计一个人类外周血基因表达和细胞组成的全面、公正的单核图谱将进行平行研究以确定细胞成分和基因表达模式。通过将神经瘤转录组与正常的图谱进行比较，手术切除了疼痛性神经瘤。神经数据集，我们计划识别疼痛神经瘤的细胞类型特异性决定因素。补充数据集为感觉神经功能障碍的分子和细胞分析提供基础疼痛综合症的研究将为疼痛的治疗选择提供见解。